Project 3 will coordinate research at BUMC with commercial research to development clinical and environmental tests for F. tularensis. As manipulation of this pathogen is potentially dangerous, rapid and accurate testing that minimizes contact with the organism is desirable. 1. Gen-Probe, Inc. will develop two diagnostic tests for F. tularensis. These are (1) a rapid DNA probe-based F. tularensis culture identification/confirmation test, and will be appropriate for general clinical laboratories and (2) a rapid amplified test for the direct detection of F. tularensis from clinical samples that will incorporate target capture and transcription medicated amplification (TMA). The detection can occur in real-time, during the amplification step. 2. Binax, Inc. will develop an easy to use point-of-care diagnostic lateral flow immunochromatographic test strip format to detect F. tularensis antigens in urine and respiratory specimens by incorporating specific antibody, produced at BUMC, to F. tularensis antigens. The test will incorporate gold colloid covalent chemistry and, if required, signal amplification methods. 3. Radiation Monitoring Devices, Inc. will design, develop and fabricate a prototype microfluidics-based instrument capable of continuous monitoring, and rapid, sensitive detection and identification of multiple aerosolized microbial pathogens. The prototype will detect and identify F. tularensis, Bacillus cereus spores, and E. coli, surrogates for B. anthracis spores and Yersinia pestis, respectively. An aerosol sampler will collect and concentrate the organisms into a continuous aqueous stream. The stream will feed into a disposable microfluidic platform where fluorescence labeling, immunocapture and fluorescence detection steps will be carried out, leading to the identification of the organisms. This prototype instrument will continuously monitor the effluent from the aerosol sampler for the presence of the three organisms for up to two hours.